Cell suspension fluidics, such as flow cytometry (FCS) and fluorescence-activated cell sorting (FACS), facilitates the identification and precise separation of individual cells based on phenotype. ...Since its introduction, flow cytometry has been used to analyze cell types and cellular processes in diverse non-vertebrate taxa, including cnidarians, molluscs, and arthropods. Ctenophores, which diverged very early from the metazoan stem lineage, have emerged as an informative clade for the study of metazoan cell type evolution. We present standardized methodologies for flow cytometry-mediated identification and analyses of cells from the model ctenophore
Mnemiopsis leidyi
that can also be applied to isolate targeted cell populations. Here we focus on the identification and isolation of ctenophore phagocytes. Implementing flow cytometry methods in ctenophores allows for fine scale analyses of fundamental cellular processes conserved broadly across animals, as well as potentially revealing novel cellular phenotypes and behaviors restricted to the ctenophore lineage.
Extracellular vesicles (EV) are purported to mediate type 2 diabetes and CVD risk and development. Physical activity and a balanced diet reduce disease risk, but no study has tested the hypothesis ...that short-term interval (INT) training would reduce EV compared with continuous (CONT) exercise in adults with prediabetes.
Eighteen obese adults (age, 63.8 ± 1.5 yr; body mass index, 31.0 ± 1.3 kg·m) were screened for prediabetes using American Diabetes Association criteria (75 g oral glucose tolerance test). Subjects were randomized to INT (n = 10, alternating 3-min intervals at 90% and 50% HRpeak, respectively) or CONT (n = 8, 70% HRpeak) training for 12 supervised sessions over 13 d for 60 min·d. Cardiorespiratory fitness (V˙ O2peak), weight (kg), as well as ad libitum dietary intake were assessed and arterial stiffness (augmentation index via applanation tonometry) was calculated using total AUC during a 75-g oral glucose tolerance test performed 24 h after the last exercise bout. Total EV, platelet EV (CD31/CD41), endothelial EV (CD105; CD31/ CD41), platelet endothelial cell adhesion molecule (PECAM) (CD31), and leukocyte EV (CD45; CD45/CD41) were analyzed via imaging flow cytometry preintervention/postintervention.
The INT exercise increased V˙O2peak (P = 0.04) compared with CONT training. Although training had no effect on platelet or leukocyte EV, INT decreased Annexin V- endothelial EV CD105 compared with CONT (P = 0.04). However, after accounting for dietary sugar intake, the intensity effect was lost (P = 0.18). Increased ad libitum dietary sugar intake after training was linked to elevated AV+ CD105 (r = 0.49, P = 0.06) and AV- CD45 (r = 0.59, P = 0.01). Nonetheless, increased V˙O2peak correlated with decreased AV+ CD105 (r = -0.60, P = 0.01).
Interval exercise training decreases endothelial-derived EV in adults with prediabetes. Although increased sugar consumption may alter EV after a short-term exercise intervention, fitness modifies EV count.
Abstract
Agrowing avenue for determining the prevalence of life beyond Earth is to search for “technosignatures” from extraterrestrial intelligences/agents. Technosignatures require significant ...energy to be visible across interstellar space and thus intentional signals might be concentrated in frequency, in time, or in space, to be found in mutually obvious places. Therefore, it could be advantageous to search for technosignatures in parts of parameter space that are mutually derivable to an observer on Earth and a distant transmitter. In this work, we used the
L
-band (1.1–1.9 GHz) receiver on the Robert C. Byrd Green Bank Telescope to perform the first technosignature search presynchronized with exoplanet transits, covering 12 Kepler systems. We used the Breakthrough Listen turboSETI pipeline to flag narrowband hits (∼3 Hz) using a maximum drift rate of ±614.4 Hz s
−1
and a signal-to-noise threshold of 5—the pipeline returned ∼3.4 × 10
5
apparently-localized features. Visual inspection by a team of citizen scientists ruled out 99.6% of them. Further analysis found two signals of interest that warrant follow up, but no technosignatures. If the signals of interest are not redetected in future work, it will imply that the 12 targets in the search are not producing transit-aligned signals from 1.1 to 1.9 GHz with transmitter powers >60 times that of the former Arecibo radar. This search debuts a range of innovative technosignature techniques: citizen science vetting of potential signals of interest, a sensitivity-aware search out to extremely high drift rates, a more flexible method of analyzing on-off cadences, and an extremely low signal-to-noise threshold.
Arterial stiffness is considered a predictor of cardiovascular disease. Females have higher values of arterial stiffness than males, suggesting a greater risk of heart-related complications. Although ...a low-calorie diet (LCD) reduces fasting arterial stiffness, in part through weight loss, it is unknown if interval exercise (INT) adds to the benefit of LCD on fasting and postprandial arterial stiffness in females with obesity.
Twenty-five females (47 ± 2.6 yr, 37.6 ± 1.3 kg·m-2) were randomized to 13 d of LCD (n = 12; mixed meals of ~1200 kcal·d-1) or LCD + INT (n = 13; 60 min·d-1 of supervised 3-min intervals at 90% HRpeak and 50% HRpeak). Arterial stiffness (augmentation index AIx and carotid-femoral pulse wave velocity cfPWV) and blood biochemistries were measured during a 75-g oral glucose tolerance test before and after the intervention to determine fasting and postprandial arterial stiffness as well as insulin sensitivity (simple index of insulin sensitivity SIIS) and inflammation (C-reactive protein, interleukin 8, and tumor necrosis factor alpha).
Although LCD + INT increased V˙O2peak and HDL compared with LCD (P = 0.04 and P < 0.01, respectively), both interventions decreased body fat, LDL, total cholesterol, and triglycerides (all P < 0.01) and increased SIIS (P = 0.03). Despite no effect on fasting AIx (P = 0.27), LCD and LCD + INT decreased AIx60min (-7.4% ± 4.3% vs -7.0% ± 5.0%, P = 0.04) and tAUC120min (-663 ± 263 vs -457 ± 406, P = 0.03). There were no changes in fasting cfPWV (P = 0.91) or cfPWV120min (P = 0.62). Increased SIIS and decreased interleukin 8 were associated with reduced fasting AIx (r = -0.44, P = 0.03, and r = 0.40, P = 0.055), whereas decreased C-reactive protein correlated with reduced postprandial AIx60min (r = 0.43, P = 0.04).
Independent of exercise, 13 d of LCD reduces postprandial AIx in females with obesity. Insulin sensitivity and inflammation correlated with improved arterial stiffness, suggesting unique mechanisms regulate fasted versus postprandial arterial stiffness.
•Caloric restriction is effective at inducing weight loss in people with obesity, although appetite hormonal responses may stimulate perceptions of feeding behavior.•Interval exercise (INT) in ...contrast may oppose desires to eat.•This study shows that a low calorie diet (LCD) with INT suppresses acylated ghrelin despite both LCD and LCD+INT stimulating peptide tyrosine-tyrosine (PYY).•LCD+INT prevented the rise in hunger observed following a LCD.•These findings suggest that performing exercise during a caloric restriction program may favor weight loss/maintenance over time in people with obesity.
Caloric restriction is suggested to increase hunger, in part, through complex interactions of hormones and behavior that contribute to challenges in long-term weight loss. Although intense exercise may attenuate appetite, no data exist testing the effects of interval exercise (INT) during a low-calorie diet (LCD) on appetite regulation. We hypothesized that LCD+INT would favorably influence satiety when compared with an energy-deficit matched LCD in women with obesity. Twenty-six women with obesity (47.3±2.4 yrs; 37.3 ± 1.2 kg/m2) were randomized to either LCD (n = 13; mixed meals of ~1200 kcal/d) or LCD+INT (n = 13; 60 min/d of supervised interval exercise at 90% HRpeak for 3 min and 50% HRpeak for 3 min) for 2 weeks. An additional 350kcal (shake) was provided to LCD+INT individuals post-exercise to equate energy availability between groups. Total PYY, acylated ghrelin and des-ghrelin were measured at 0, 30 and 60 min of a 75g OGTT before and after the intervention. Visual analog scales were also administered at 0 and 120 min of the OGTT to assess appetite perception. Food logs were recorded prior to and during the intervention to ensure caloric intake compliance. Compared with pre-intervention conditions, both interventions decreased food intake (P = 0.001) and body fat (P < 0.01). There was no effect on fasting PYY, but both LCD and LCD+INT increased post-prandial PYY iAUC (P < 0.001) relative to pre-intervention. LCD+INT maintained fasting acylated ghrelin (P = 0.06) and suppressed post-prandial acylated ghrelin iAUC (P = 0.04) compared to LCD. Neither intervention impacted circulating des- ghrelin before or following the OGTT. Interestingly, LCD+INT attenuated fasting hunger and maintained fullness compared with LCD (P = 0.05 and P = 0.06, respectively). Taken together, interval exercise favors acylated ghrelin suppression and perception of hunger during a LCD in women with obesity.
The effect of work-matched exercise intensity on β-cell function is unknown in people with prediabetes before clinical weight loss. We determined if short-term moderate continuous (CONT) vs. ...high-intensity interval (INT) exercise increased β-cell function. Thirty-one subjects (age: 61.4 ± 2.5 yr; body mass index: 32.1 ± 1.0 kg/m
) with prediabetes American Diabetes Association criteria, 75-g oral glucose tolerance test (OGTT) were randomized to work-matched CONT (70% HR
) or INT (3 min 90% HR
and 3 min 50% HR
) exercise for 60 min/day over 2 wk. A 75-g 2-h OGTT was conducted after an overnight fast, and plasma glucose, insulin, C-peptide, and free fatty acids were determined for calculations of skeletal muscle oral minimal model (OMM), hepatic (homeostatic model of insulin resistance), and adipose (Adipose-IR) insulin sensitivity. β-Cell function was defined from glucose-stimulated insulin secretion (GSIS, deconvolution modeling) and the disposition index (DI). Glucagon-like polypeptide-1 GLP-1(active) and glucose-dependent insulinotropic polypeptide (GIP) were also measured during the OGTT, along with peak oxygen consumption and body composition. CONT and INT increased skeletal muscle- but not hepatic- or adipose-derived DI ( P < 0.05). Although both treatments tended to reduce fasting GLP-1(active) ( P = 0.08), early phase GLP-1(active) increased post-CONT and INT training ( P < 0.001). Interestingly, CONT exercise increased fasting GIP compared with decreases in INT ( P = 0.02). Early and total-phase skeletal muscle DI correlated with decreased total glucose area under the curve ( r = -0.52, P = 0.002 and r = -0.50, P = 0.003, respectively). Independent of intensity, short-term training increased pancreatic function adjusted to skeletal muscle in relation to improved glucose tolerance in adults with prediabetes. Exercise also uniquely affected GIP and GLP-1(active). Further work is needed to elucidate the dose-dependent mechanism(s) by which exercise impacts glycemia. NEW & NOTEWORTHY Exercise is cornerstone for reducing blood glucose, but whether high-intensity interval training is better than moderate continuous exercise is unclear in people with prediabetes before weight loss. We show that 2 wk of exercise training, independent of intensity, increased pancreatic function in relation to elevated glucagon-like polypeptide-1 secretion. Furthermore, β-cell function, but not insulin sensitivity, was also correlated with improved glucose tolerance. These data suggest that β-cell function is a strong predictor of glycemia regardless of exercise intensity.
The optimal short-term exercise dose to improve glucose tolerance in relation to metabolic flexibility and/or insulin resistance is unknown. Therefore, we tested if short-term, work-matched ...continuous (CONT) versus interval (INT) exercise training improves glucose tolerance in part by reducing insulin resistance and increasing metabolic flexibility independent of clinically meaningful fat loss in adults with prediabetes.
Subjects (age = 60.9 ± 1.4 yr, body mass index = 33.5 ± 1.1 kg·m) were screened for prediabetes using the American Diabetes Association criteria (75 g oral glucose tolerance test OGTT and/or HbA1c) and were randomized to 60 min·d of supervised CONT (n = 17, 70% HRpeak) or work-matched INT (n = 14; 90% HRpeak for 3 min and 50% HRpeak for 3 min) exercise for 12 bouts. Fitness (V˙O2peak) and body composition were assessed pre- and postintervention. A 180-min 75-g OGTT was performed, and glucose, insulin, and free fatty acids were collected to calculate glucose tolerance (tAUC180min) and whole-body as well as adipose tissue insulin resistance pre- and postintervention. RER (indirect calorimetry) was also measured at 0, 60, 120, and 180 min of the OGTT to assess fasting and postprandial metabolic flexibility.
CONT and INT training improved V˙O2peak (L·min; P = 0.001) and glucose tolerance (P = 0.01) and reduced fasting RER (P = 0.006), as well as whole-body and adipose insulin resistance (both P = 0.02) with no effect on body fat (P = 0.18). Increased postprandial RER was correlated with reduced glucose tAUC180min (r = -0.38, P = 0.05) and increased 180-min RER related to decreased whole-body insulin resistance (r = -0.42, P = 0.03).
Independent of exercise dose and fat loss, short-term training improves glucose tolerance in relation to enhanced postprandial fuel use.
Repetitive negative thinking (RNT) is a key transdiagnostic mechanism underpinning depression and anxiety. Using "just-in-time adaptive interventions" via smartphones may disrupt RNT in real time, ...providing targeted and personalized intervention.
This pilot randomized controlled trial evaluates the feasibility, acceptability, and preliminary clinical outcomes and mechanisms of Mello-a fully automated, personalized, transdiagnostic, and mechanistic smartphone intervention targeting RNT in young people with depression and anxiety.
Participants with heightened depression, anxiety, and RNT were recruited via social media and randomized to receive Mello or a nonactive control over a 6-week intervention period. Assessments were completed via Zoom sessions at baseline and at 3 and 6 weeks after baseline.
The findings supported feasibility and acceptability, with high rates of recruitment (N=55), uptake (55/64, 86% of eligible participants), and retention (52/55, 95% at 6 weeks). Engagement was high, with 90% (26/29) and 59% (17/29) of the participants in the Mello condition still using the app during the third and sixth weeks, respectively. Greater reductions in depression (Cohen d=0.50), anxiety (Cohen d=0.61), and RNT (Cohen d=0.87) were observed for Mello users versus controls. Mediation analyses suggested that changes in depression and anxiety were accounted for by changes in RNT.
The results indicate that mechanistic, targeted, and real-time technology-based solutions may provide scalable and effective interventions that advance the treatment of youth mental ill health.
Australian New Zealand Clinical Trials Registry ACTRN12621001701819; http://tinyurl.com/4d3jfj9f.
No short-term exercise data exist testing whether training intensity modifies hormonal and perceived appetite in obese adults with prediabetes. Therefore, we compared the effects of short-term ...moderate-continuous (CONT) vs. high-intensity interval (INT) training on appetite regulation. Twenty-eight obese adults age: 61.3 ± 1.5 yr; body mass index (BMI): 33.2 ± 1.1 kg/m
with prediabetes were randomized to work-matched CONT ( n = 14) or INT ( n = 14) training for 2 wk. Plasma acylated ghrelin (AG), des-acylated ghrelin (dAG), active glucagon-like peptide-1 (GLP-1), and insulin were measured at 0, 30, and 60 min of a 75-g oral glucose tolerance test (OGTT) before and after training. Visual analog scales were administered at 0 and 120 min during the OGTT to examine perceived appetite. Three-day food logs were collected before and after testing to assess ad libitum diet. CONT and INT increased peak oxygen consumption ( P < 0.01) and decreased BMI ( P < 0.01). Although neither intervention altered fasting levels of AG ( P = 0.94), dAG ( P = 0.36), or insulin ( P = 0.67), CONT raised GLP-1 compared with INT ( P = 0.05). Exercise training did not affect postprandial suppression of AG ( P = 0.81) and dAG ( P = 0.67) or stimulation of GLP-1 ( P = 0.67) and insulin ( P = 0.32). Both interventions tended to decrease total energy and protein intake ( P = 0.09 and P = 0.05, respectively), despite no change in fasting hunger ( P = 0.88) and reduced perceived fullness at 120 min during the OGTT ( P = 0.05). We conclude that 2 wk of exercise training intensity does not modulate appetite-regulatory hormones in obese adults with prediabetes. Although perceived fullness to the OGTT was reduced after exercise, CONT and INT decreased energy intake, suggesting that exercise does not elicit compensatory appetite behavior to gain weight. NEW & NOTEWORTHY Adults with prediabetes are at risk for appetite dysregulation. Although exercise promotes weight management, it is unclear whether moderate-continuous or high-intensity interval training is more beneficial for appetite regulation. We show that 2 wk of exercise, independent of intensity, does not alter postprandial appetite hormones or hunger, despite slight reductions in food intake and weight. These data support exercise as an effective method to induce negative energy balance without compensatory weight gain.
A segmented genome enables influenza virus to undergo reassortment when two viruses infect the same cell. Although reassortment is involved in the creation of pandemic influenza strains and is ...routinely used to produce influenza vaccines, our understanding of the factors that drive the emergence of dominant gene constellations during this process is incomplete. Recently, we defined a spectrum of interactions between the gene segments of the A/Udorn/307/72 (H3N2) (Udorn) strain that occur within virus particles, a major interaction being between the NA and PB1 gene segments. In addition, we showed that the Udorn PB1 is preferentially incorporated into reassortant viruses that express the Udorn NA. Here we use an influenza vaccine seed production model where eggs are coinfected with Udorn and the high yielding A/Puerto Rico/8/34 (H1N1) (PR8) virus and track viral genotypes through the reassortment process under antibody selective pressure to determine the impact of Udorn NA-PB1 co-selection. We discovered that 86% of the reassortants contained the PB1 from the Udorn parent after the initial co-infection and this bias towards Udorn PB1 was maintained after two further passages. Included in these were certain gene constellations containing Udorn HA, NA, and PB1 that confered low replicative fitness yet rapidly became dominant at the expense of more fit progeny, even when co-infection ratios of the two viruses favoured PR8. Fitness was not compromised, however, in the corresponding reassortants that also contained Udorn NP. Of particular note is the observation that relatively unfit reassortants could still fulfil the role of vaccine seed candidates as they provided high haemagglutinin (HA) antigen yields through co-production of non-infectious particles and/or by more HA molecules per virion. Our data illustrate the dynamics and complexity of reassortment and highlight how major gene segment interactions formed during packaging, in addition to antibody pressure, initially restrict the reassortant viruses that are formed.
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